Chen Xiaotao, Cui Yubao, Ma Yanming
Department of Orthopedics, Qinghai Provincial People's Hospital, Xining City, Qinghai Province 810007, China.
Department of Orthopadics, Hubei Aerospace Hospital, Xiaogan City, Hubei Province 432000, China.
J Bone Oncol. 2020 Aug 2;25:100314. doi: 10.1016/j.jbo.2020.100314. eCollection 2020 Dec.
Osteosarcoma is the most common type of bone malignancy. Increasing evidence indicated that long non-coding RNAs (lncRNAs) possess multiple functions in the development of cancer and can be used as indicators of prognosis and diagnosis. LncRNA BLACAT1 has been found to promote the proliferation of breast cancer cells. However, the role of BLACAT1 in osteosarcoma remains largely unknown.
QRT-PCR analysis was employed to evaluate mRNA expressions. Western blot was performed to measure relevant protein level. Colony formation and EdU assays were conducted to certify proliferative ability. TUNEL assay was finalized to assess apoptotic cells. Wound-healing and transwell assays were utilized for the exploration of migrating and invasive abilities. The subcellular distribution of BLACAT1 was studied by nucleus-cytoplasm separation assay. Relevant mechanical experiments were combined to elucidate molecular relationship between molecules.
BLACAT1 was highly expressed in osteosarcoma. BLACAT1 promoted the proliferation and migration of osteosarcoma cells. BLACAT1 acted as a sponge for miR-608 to augment the expression of Sex determining region Y-box protein 12 (SOX12), the direct target of miR-608. Further, inhibiting miR-608 recovered the repressive effect of silenced BLACAT1 on the malignant behaviors of osteosarcoma cells.
This study highlighted the contribution of BLACAT1/miR-608/SOX12 axis to the progression of osteosarcoma, suggesting novel targets for osteosarcoma therapy.
骨肉瘤是最常见的骨恶性肿瘤类型。越来越多的证据表明，长链非编码RNA（lncRNA）在癌症发展中具有多种功能，可作为预后和诊断指标。已发现lncRNA BLACAT1可促进乳腺癌细胞增殖。然而，BLACAT1在骨肉瘤中的作用仍 largely未知。
采用QRT-PCR分析评估mRNA表达。进行蛋白质印迹法检测相关蛋白水平。进行集落形成和EdU试验以验证增殖能力。完成TUNEL试验以评估凋亡细胞。利用伤口愈合试验和transwell试验探索迁移和侵袭能力。通过核质分离试验研究BLACAT1的亚细胞分布。结合相关力学实验阐明分子间的分子关系。
BLACAT1在骨肉瘤中高表达。BLACAT1促进骨肉瘤细胞的增殖和迁移。BLACAT1作为miR-608的海绵，增强了miR-608的直接靶标性别决定区Y盒蛋白12（SOX12）的表达。此外，抑制miR-608可恢复沉默的BLACAT1对骨肉瘤细胞恶性行为的抑制作用。
本研究突出了BLACAT1/miR-608/SOX12轴对骨肉瘤进展的作用，为骨肉瘤治疗提供了新的靶点。
